Genetic Diversity Of Paconia Rockii Revealed By ISSRs

Paeonia rockii blongs to Paeonia section moutan DC,it wes named because the base of every petals have a obvious stain.It has high ornamental value,because of its beautiful flowers. It also had relatively wide range of ecological adaptability, cold resistance, drought tolerance, humidity tolerance, disease resistance and strong resistance barren and other excellent properties.They are valuable resources to the peony breeding and improvement to genetic traits of. The genetic diversity of a population in6of Paeonia rockii material was analyzed using inter-simple sequence repeat markers (ISSR). Fifteen primers selected from100primers were used for ISSR amplification.Draw the following main conclusions:1. The optimal ISSR reaction system and reaction conditions of Paeonia rockii were finally established:25μL total volume of PCR amplification, included0.7μL (50ng) template DNA,10×PCR buffer2.2μL,1.2mmol·L-1MgC12,1.2mmol·L-1dNTPs1μL,1.0μmol·L-1primer2μL,5U/μL Tap DNA polymerase0.3μL, supplement distill water.Reaction condition:94℃3min,94℃degeneration45s,72℃extension90s,35cycles,72℃extension7min.2. Fifteen primers selected from100primers were used for ISSR amplification. A total of134bands were generated, of which96bands were polymorphic bands (the percentage of polymorphic band, PPB=71.6%). The molecular weight of the fragment DNA between100and2000bp. the highest percentage of polymorphism were808and844, reached85.7%. The minimum was50%blonged to827.3. According to the results of ISSR amplification, which were analyzed to Jaccard similarity coefficient by NTSYSpc2.10e software, the genetic similarity coefficient was0.45to0.93.The clustering dendrogram was constructed by UPGMA method. Sixteen P.rockii materials were divided into four major groups while the similarity coefficient was0.534. It was can been saw By clustering graph, in the16of Paeonia rockii, P. rockii ’shushengpengmo’ and P. rockii ’xuehaibingxin’ hold the most nearest genetic similarity coefficient,it is0.93. But the farthest genetic similarity coefficient is0.45, between P. rockii ’fenpantuogui’ and P. rockii ’antangjingxin’4. Based on the NTSYSpc2.10e software The clustering dendrogram was constructed by UPGMA method. Sixteen P.rockii materials were divided into four major groups:Class Ⅰ included six of Paeonia rockii materials, and five in six was crown form, left a single form species. But other classes also have crown flower varieties. Class Ⅱ had a anemone form specie, it was the only anemone species elected in the selected materials.Four different color and form materials were clustered into Cass Ⅲ, suggest that the character of closer phylogenetic relationship species were varieties. A group of Class Ⅳ included four white flowers materials, whose form were diffrent. Group B was P. rockii ’yinxiannv’ alone, it was the only red material selected. Thus, genetic clustering classification of different resemble coefficients was not consisted with that based on the flower color and form.